Nonanedioic acid, 1,9-bis(2-octyldodecyl) ester

Genetic toxicity: in vivo

Administrative data

Endpoint:

in vivo mammalian somatic cell study: cytogenicity / erythrocyte micronucleus

Remarks:

Type of genotoxicity: chromosome aberration

Type of information:

migrated information: read-across based on grouping of substances (category approach)

Adequacy of study:

key study

Study period:

11 Nov 1985 - 28 Jan 1986

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Acceptable, well documented study report which meets basic scientific priciples. Purity of test substance not given.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1986

Report date:

1986

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 474 (Mammalian Erythrocyte Micronucleus Test)

Version / remarks:

adopted in 1981

Deviations:

yes

Remarks:

purity of test substance not given

GLP compliance:

yes

Type of assay:

micronucleus assay

Test material

Test material information

Constituent 1

Reference substance name:

69275-01-0

Cas Number:

69275-01-0

IUPAC Name:

69275-01-0

Details on test material:

- Name of test material (as cited in study report): Di-(2-octyl-dodecyl)-sebacate
- Physical state: colourless, viscous liquid
- Analytical purity: no data

Test animals

Species:

mouse

Strain:

other: CFW 1

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Winkelmann
- Age at study initiation: 7 weeks
- Weight at study initiation: 18 - 31g
- Assigned to test groups randomly: yes
- Fasting period before study: 4 hours for 10000mg/kg bw group, 6-16 hours for other test groups
- Housing: one animal per cage in Makrolon Type I cages (male), 4 animals per cage in Makrolon Type II cages (female)
- Diet: Altromin 1324 diet, ad libitum
- Water: tap water, ad libitum

ENVIRONMENTAL CONDITIONS
- Photoperiod (hrs dark / hrs light): 12/12

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

- Vehicle(s)/solvent(s) used: oleum arachidis G 166, DAB8
- Amount of vehicle (if gavage or dermal): 10mL/kg bw

Details on exposure:

PREPARATION OF DOSING SOLUTIONS:
test substance was diluted using Oleum Arachidis G 166, DAB8 before application by gavage

Duration of treatment / exposure:

negative control, positive control, 1000 & 5000 mg/kg bw: 24 hours
10000 mg/kg bw: 24, 48, 72 hours

Frequency of treatment:

single treatment

Post exposure period:

24, 48 and 72 hours

Doses / concentrations

Remarks:

Doses / Concentrations:
1000, 5000, 10000 mg/kg bw
Basis:
actual ingested

No. of animals per sex per dose:

7

Control animals:

yes, concurrent vehicle

Positive control(s):

cyclophosphamide
- Route of administration: intraperitoneal
- Doses / concentrations: 10 mg/kg bw

Examinations

Tissues and cell types examined:

tissue: bone marrow from the femur
cell type: bone marrow cells

Details of tissue and slide preparation:

CRITERIA FOR DOSE SELECTION: Range finding study performed to find the maximum tolerated dose.

TREATMENT AND SAMPLING TIMES (in addition to information in specific fields): 24, 48 and 72 hours

DETAILS OF SLIDE PREPARATION: 3 slides per animal were fixed with methanol, stained for 10 minutes with Giemsa-solution (modified from Gollapudi and Kamra, 1979), rigorously washed with bidest water and air-dried over night.

METHOD OF ANALYSIS: 1 slide per animal was analyzed. The number of micronuclei in 1000 polychromatic erythrocytes was counted, additionally the ratio polychromatic erythrocytes to normocytes was determined.

Evaluation criteria:

Only regions on slides with a good cell distribution, intact cells and sufficient staining were evaluated. The ratio of polychromatic to normochromatic erythrocytes was determined by evaluating the number of bluish to red stained cells.

Statistics:

The statistic significance was calculated in comparison to controls using tables by Kastenbaum and Bowman (Mutation Research 2: 527-549; 1970).

Results and discussion

Test results

Sex:

male/female

Genotoxicity:

negative

Toxicity:

yes

Remarks:

5000, 10000 mg/kg bw dose groups: male and female animals showed slightly reduced activity and ruffled fur. Symptoms had disappeared 20 hours after application.

Vehicle controls validity:

valid

Negative controls validity:

valid

Positive controls validity:

valid

Additional information on results:

RESULTS OF RANGE-FINDING STUDY
- Dose range: 5000 - 10000 mg/kg bw
- Clinical signs of toxicity in test animals: none

Any other information on results incl. tables

Table 1: Results of the in vivo micronucleus assay in male animals.
			Total micronuclei per 1000 PCEs at sampling time			Mean PCEs / 1000 NCEs at sampling time
Exp group	Number of animals	Dose [mg/kg bw]	24 h	48 h	72 h	24 h	48 h	72 h
Vehicle control (oleum Arachidis, DAB8), 24 h	7	10 mL/kg bw	2	n.d.	n.d.	0.73	n.d.	n.d.
Positive control (cyclophosphamide), 24 h	7	10	8.71*	n.d.	n.d.	0.91	n.d.	n.d.
Test substance	7	1000	n.d.	n.d.	n.d.	n.d.	n.d.	n.d.
Test substance	7	5000	n.d.	n.d.	n.d.	n.d.	n.d.	n.d.
Test substance	7	10000	0.86	1.57	1.71	0.94	1.11	1.07
*statistically significant (p>0.01), n.d. = not determined; in the highest dose group (10000 mg/kg bw) the group mean micronucleated cell count was comparable with the concurrent control values, therefore dosages of 1000 and 5000 mg/kg bw have not been evaluated.
Table 2: Results of the in vivo micronucleus assay in female animals.
			Total micronuclei per 1000 PCEs at sampling time			Mean PCEs / 1000 NCEs at sampling time
Exp group	Number of animals	Dose [mg/kg bw]	24 h	48 h	72 h	24 h	48 h	72 h
Vehicle control (oleum Arachidis, DAB8), 24 h	7	10 mL/kg bw	2.14	n.d.	n.d.	0.95	n.d.	n.d.
Positive control (cyclophosphamide), 24 h	7	10	8.57*	n.d.	n.d.	0.88	n.d.	n.d.
Test substance	7	1000	n.d.	n.d.	n.d.	n.d.	n.d.	n.d.
Test substance	7	5000	n.d.	n.d.	n.d.	n.d.	n.d.	n.d.
Test substance	7	10000	1.14	1.29	1.71	0.91	1.10	1.18
*statistically significant (p>0.01), n.d. = not determined; in the highest dose group (10000 mg/kg bw) the group mean micronucleated cell count was comparable with the concurrent control values, therefore dosages of 1000 and 5000 mg/kg bw have not been evaluated.

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information): negative